Dietary fish oil replacement by soybean oil: Effect on plasma vitellogenin,sex steroids and ovarian steroidogenesis in Chinese strip‐necked turtles (Mauremys sinensis)

In order to investigate the effects of dietary fish oil replacement, the turtles (Mauremys sinensis) were fed four experimental diets for 10 months: FO (100% fish oil), FSO (70% fish oil and 30% soybean oil), SFO (30% fish oil and 70% soybean oil) and SO (100% soybean oil), sampled at pre‐vitellogenesis, vitellogenesis and post‐vitellogenesis. The results showed that plasma gonadotropin‐releasing hormone (GnRH) levels were the highest at pre‐vitellogenesis, which promoted the secretion of gonadotropin and sex steroids. Therefore, plasma luteinizing hormone (LH) and estrogen (E₂) levels were significantly increased at post‐vitellogenesis (p < 0.05), while follicle‐stimulating hormone (FSH) levels increased at vitellogenesis (p < 0.05). The FO and FSO groups had significantly higher GnRH and E₂ levels than the other two groups (p < 0.05). In addition, plasma vitellogenin (Vtg) levels significantly increased at vitellogenesis and post‐vitellogenesis (p < 0.05), which were significantly higher in the groups of FO and FSO than SO (p < 0.05). Moreover, the expression levels of hepatic estrogen receptor α (Erα) mRNA were significantly increased at vitellogenesis and post‐vitellogenesis while ovarian Cyp19α1α mRNA were significantly increased at post‐vitellogenesis (p < 0.05), and both were the lowest in SO. Taken together, the replacement of fish oil with 66.7% soybean oil is feasible.     

The relationship between plasma and ovarian levels of gonadal steroids in the repeat spawning marine fishesPagrus auratus (Sparidae) andChromis dispilus (Pomacentridae)

The relationship between plasma and ovarian levels of gonadal steroids was examined in two New Zealand fish species with multiple spawning cycles of differing length. Snapper (Pagrus auratus) have a daily cycle of oocyte development, ovulation and spawning, whereas demoiselles (Chromis dispilus) spawn over 2–3 days during a repeat spawning cycle of 7–9 days. Ovarian and plasma levels of the gonadal steroids 17β-estradiol (E₂), testosterone (T), 17-hydroxyprogesterone (17P) and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) were measured in reproductively active fish captured from the wild. Ovarian levels of E₂, T and 17P changed in relation to spawning cycle and gonad stage in both snapper and demoiselles. E₂ and T levels were detectable at all times, but highest during vitellogenesis in both species. Cyclic changes of 17P occurred in both species, and levels appeared to depend on the rate of conversion of 17P to other hormones. No changes in ovarian levels of 17,20βP were detected in relation to stage of the spawning cycle in snapper; however, ovarian levels of 17,20βP were highest in demoiselles before spawning when fish undergoing final oocyte maturation predominated. Plasma levels of E₂ and T were strongly correlated with ovarian concentrations (r=0.850 and r=0.819 for E₂ and T respectively) in demoiselles but there was poor correlation between ovarian and plasma levels of 17P and 17,20βP (r=0.004 and 0.273 respectively), or between ovarian and plasma levels of E₂, T, 17P or 17,20βP of snapper (r=0.135, 0.277, 0.131 and 0.279). The poor correlation between plasma and ovarian levels of some steroid hormones suggests that plasma concentrations of steroids may not adequately reflect the reproductive status of the fish during short-term cyclic ovarian changes. It is suggested that this disparity is likely to be most marked in species with ovulatory periodicity of short duration.     

Estradiol-17β suppresses testicular development and stimulates sex reversal in protandrous black porgy,Acanthopagrus schlegeli

Two year old black porgy (Acanthopagrus schlegeli) fed a diet containing 4.0 mg kg^–1 of estradiol-17 (E₂) for 5 months had significantly lower GSI than the control group during the spawning season. E₂ suppressed testicular development, spermiation and plasma testosterone (T) and 11-ketotestosterone (11-KT) and stimulated ovarian development, vitellogenesis and sex reversal. Spermiation in the control group occurred in January and February with the concentrations of 1.08–1.36 × 10¹⁰ sperm ml^–1 of milt. Higher plasma T and 11-KT, but lower E₂ levels were detected in the spermiating fish (control group). Higher plasma E₂ levels were detected in the sex reversing black porgy during the pre-spawning season. A sharp rise in plasma 11-KT and a drop in T levels were detected in spermiating fish (control group) from January to February. Plasma 11-KT levels correlated with the testicular development and spermiation. The data suggest that E₂ plays an important role in controlling the sex reversal of black porgy.to whom correspondence should be addressed.     

Gonadal histology and some biochemical characteristics of <Emphasis Type="Italic">Chalcalburnus tarichi</Emphasis> (Pallas, 1811) having abnormal gonads

The gonad histology, gonado-somatic index (GSI), 17β-estradiol (E₂) levels and acetylcholinesterase (AChE) activity in the carp species Chalcalburnus tarichi from Lake Van and the Karasu river, eastern Turkey, have been investigated. Fish between 5 and 7 years old were sampled from November 2003 to February 2004. The ratio of female fish caught in Lake Van with abnormal ovaries (AbOF) was 43.3%, but the fork length and body weight of these fish were not correlated with this abnormality. The weight of the ovaries and the GSI values of AbOF were very low (P < 0.05). Histological observations on the samples caught each month revealed that the oocytes had degenerated in the perinucleolus and early cortical alveolus stages and that the ovaries were full of somatic stromal tissue. In addition, the seminiferous tubules of male fish with abnormal testes did not contain male reproductive cells at any stage. The ovaries of the fish caught from the Karasu river were also full of oocytes in the perinucleolus and early cortical alveolus stages, but there were fewer atretic follicles. Furthermore, apoptosis was observed in the ovary cells of these fish, in particular in the follicular cells, and the plasma E₂ levels of the AbOF was very low (P < 0.05). AChE activity was inhibited only in liver (P < 0.05). We conclude that our sample of C. tarichi must have been exposed to various polluting chemicals or another unknown factors (such as global warming) and that these factors have irreversibly impaired oocyte development in a high percentage of fish.     

Influence of 11-ketotestosterone, 17β-estradiol, and 3,5,3′-triiodo-L-thyronine on distribution and metabolism of carotenoids in Arctic charr, Salvelinus alpinus L.*

This investigation examines the influence of implants containing 11-ketotestosterone (11KT), 17-estradiol (E₂), and 3,5,3-triiodo-l-thyronine (T₃) on astaxanthin metabolism in sexually immature individually tagged Arctic charr. The fish (initial average weight 427 g) were maintained in freshwater for 40 days, and weekly implanted intraperitoneally with oil-based injections containing either 11 KT, E₂ or T₃ at levels of 0.1, 1.0 and 0.1 mg (100 g body weight (BW))^–1, respectively. The control fish were given the oil medium alone (0.2 ml 100 g BW^–1). The diet contained ca. 50 mg astaxanthin kg^–1. Carotenoid composition was monitored in plasma, fillet, liver and skin, and 11 KT, E₂ and testosterone (T) levels in plasma. All hormone treatments reduced plasma T compared to the control. E₂-treated fish had a higher (p<0.05) hepatosomatic index (HSI) than the other treatments. Hormone treatment did not influence gonadosomatic index (GSI). T₃ administration induced a silvery skin appearance. The fillet and plasma carotenoid content decreased during the experiment. 11 KT implantation reduced astaxanthin and idoxanthin concentrations of plasma and fillets, and increased the amount in liver and skin, compared to the other treatments. The relative proportion of astaxanthin to idoxanthin was higher in the control fish and T₃ implanted fish, than in fish implanted with 11 KT or E₂ (p<0.05). Fish treated with E₂ had the highest skin carotenoid concentration. Male fish had significantly higher carotenoid content in plasma, fillet and skin than female fish. This study reveals that sex hormones affect carotenoid metabolism and partitioning among body compartments of Arctic charr, effects differently displayed by the sexes.     

Estradiol-17β-induced calcium uptake and resorption in juvenile rainbow trout,Oncorhynchus mykiss

Juvenile rainbow trout, Oncorhynchus mykiss, were injected with estradiol-17β (E₂) in order to study the source of extra calcium needed during vitellogenesis. E₂-treatment increased the calcium uptake from the external medium as well as calcium mobilization from muscle and scale. Judged by the increase in plasma protein-bound calcium levels, the E₂-induced increase in calcium uptake is an apparent over-mobilization of calcium, i.e., the calcium uptake of the fish is in excess of what is found bound to plasma proteins. As the calcium excretion and calcium space (calculated from free plasma calcium levels) were unaffected, the excess calcium is suggested to be incorporated into internal calcium stores. This implies that the systems regulating vitellogenesis and calcium balance are integrated on the mechanistic or endocrine level, and that E₂ causes calcium mobilization of a magnitude geared to the needs of the sexually maturing female.     

Ovarian re‐maturation following the first spawning in the Chinese mitten crab,Eriocheir sinensis (H. Milne‐Edwards)

The present study investigated the external and histological changes in the ovary and measured the gonadosomatic index (GSI) and hepatosomatic index (HSI) during the re‐maturation of the Chinese mitten crab Eriocheir sinensis. The results show that the ovarian re‐maturation cycle of this crab species can be divided into four stages, i.e. Stage I: the ovary had an X shape and appeared milky white or with a mottled buff colour, and the dominant gametocyte at the stage were oogonia (OG, 28.3%) and previtellogenic oocytes (PR, 31.6%); Stage II: the ovary appeared light yellow or tan in colour, dominated by endogenous vitellogeic oocytes (EN, 69.3%). The yolk globules of unreleased mature oocytes (MO) from the previous spawn were absorbed, leaving many vacuoles in the retrogressing MO; Stage III: the ovary appeared crimson red or deep purple, and although the main type of gametocyte was exogenous vitellogenic oocytes (EX), near‐mature oocytes (NO) could also be found in the late phase of the ovarian development stage; Stage IV: the ovary was ripe and filled with MO that contained large yolk globules. During the period of the second ovarian maturation cycle, the GSI increased significantly from the time just after the first spawning (P<0.01). However, although HSI appeared to decrease within days after spawning (DAS), no significant correlation was found between GSI and HSI or between HSI and DAS (P>0.05). Furthermore, the final GSI and the volume of MO of E. sinensis during the second maturation cycle were drastically lower than those of the first maturation cycle.     

Biological properties of Indian walking catfish (Clarias batrachus) (L.) gonadotropins in female reproduction

The biological activities of catfish LH-like (semi-purified: s200a and purified Qa) and FSH-like (semi-purified: s200b and purified: Qb) were compared in intact and hypophysectomized female catfish, Clarias batrachus, during preparatory and the pre-spawning periods on vitellogenesis and ovarian maintenance, as well as in vitro final maturation of oocytes, germinal vesicle breakdown (GVBD). During preparatory period, in intact catfish, semi-purified FSH-like induced complete vitellogenesis through the production of estradiol-17β (E₂) and vitellogenin (Vg) accompanied by the formation of SIII yolky oocytes. On the other hand, semi-purified LH-like had induced the formation of only SII (characterized by the appearance of cortical alveoli in cytoplasm) oocytes, which indicates the initiation of vitellogenesis. In hypophysectomized female catfish, purified LH-like but not FSH-like induced the formation of SII oocytes in the ovaries. Treatment with semi-purified LH- and FSH-like at the dose level of 5 µg/fish/day for 7 days significantly maintained the yolky oocytes in gravid catfish after hypophysectomy with a significant reduction in plasma Vg, but not E₂ levels, indicating some unknown GtH-induced factor doing the job. In in vitro oocytes culture, both LH- and FSH-like induced GVBD, but the response was significantly more with LH-like than FSH-like. All these findings revealed that both LH-like and FSH-like have overlapping physiological functions, but their responses differ depending on the physiological status of the catfish.     

Purification,characterization and immunoassay of striped bass (Morone saxatilis) vitellogenin

The egg yolk precursor, vitellogenin (VTG), was purified from blood plasma of striped bass by chromatography on hydroxylapatite or DEAE-agarose. The fish were first implanted with estradiol-17β (E₂), which induced vitellogenesis. A rabbit antiserum (a-FSPP) raised against plasma from mature female striped bass, and then adsorbed with mature male plasma, was used to detect female-specific plasma protein (FSPP) in the chromatography fractions. Striped bass VTG (s-VTG) was collected from the peak fraction that was induced by E₂, reacted with a-FSPP, and contained all detectable phosphoprotein. It appeared as a single band (M_r ≂ 170,000) in SDS-PAGE or Western blots using a-FSPP, and as a pair of closely-spaced phospholipoprotein bands in native gradient-PAGE, suggesting that there is more than one circulating form of s-VTG. The relationship of s-VTG to the yolk proteins was verified using a-FSPP. The antiserum reacted with the main peak from gel filtration of saline ovary extracts, and it specifically immunostained the two main bands in Western blots of the extracts and the yolk granules of mature oocytes. The amino acid composition of s-VTG was similar to that of VTG from other fish and Xenopus. A radial immunodiffusion assay for s-VTG was developed using a-FSPP and purified s-VTG as standard. The s-VTG was not detected in blood plasma of males, immature females, or regressed adult females, but plasma s-VTG levels were highly correlated with plasma E₂ and testosterone levels, and oocyte growth, in maturing females. The results indicate that the maturational status of female striped bass can be identified by s-VTG immunoassay.     

Correlation between oocyte development and plasma concentrations of steroids and vitellogenin in greenback flounder Rhombosolea tapirina

The development of oocytes in association with changes in plasma concentrations of vitellogenin (Vtg), 17β-estradiol (E₂) and testosterone (T) was investigated in maturing female greenback flounder Rhombosolea tapirina over the first part of a reproductive season. During vitellogenesis, increasing oocyte size was accompanied by the sustained elevation of plasma Vtg and E₂. There was a marked increase in T towards the end of vitellogenesis.     

Effects of Thyroxin and Domperidone on Oocyte Maturation and Spawning Performances in the Rabbit Fish,Siganus guttatus

Thyroxin (T₄) is proven to have various functions in fish reproduction. In this study, we investigated oocyte maturation and spawning performance in the rabbit fish, Siganus guttatus, orally exposed to different doses of T₄ (0.05, 0.25, and 0.5 mg/kg fish) during vitellogenesis. No statistically significant differences of reproductive characteristics were observed among the treatments. However, the group that was exposed to the dose of 0.5 mg/kg spawned after receiving a hormone injection. The findings suggest that feeding T₄ during vitellogenesis did not advance oocyte maturation, but probably could induce spawning in rabbit fish. In a separate experiment, we investigated the effects of orally administered domperidone (DOM) on reproductive performances of the female rabbit fish. The study showed that oral administration of DOM during the early stages of the reproductive cycle had no significant effects on oocyte maturation and spawning performance. Overall, these findings suggest that DOM may not be needed for the induction of maturation and spawning of this species under aquaculture conditions. These practical investigations were conducted with a view to attempt reducing drug use, production costs, and support sustainable aquaculture.     

The reproductive physiology of three age classes of adult female diploid and triploid brook trout (Salvelinus fontinalis)

Triploid female fish show impaired gametogenesis and are unable to produce viable offspring. The reproductive physiology of artificially-induced triploid female salmonids has been well described up until the time of first sexual maturation in diploids, but few reports exist for older triploids. This study reports the influence of triploidy on growth, ovarian development and reproductive endocrinology among three age classes of female brook trout (Salvelinus fontinalis) in comparison to sibling diploids. Triploids were larger than diploids for most of the study period, but the difference was statistically significant only during maturation and spawning of 2⁺ diploids. Plasma estradiol-17 (E₂), testosterone (T) and vitellogenin (VTG) levels in triploids were generally lower than in diploids, and VTG was the only parameter to show seasonal fluctuations resembling those of diploids. Triploids showed significantly lower GSI and total oocyte number than diploids of similar age, and only half of all triploids sacrificed during the study (n=56) had developing oocytes in their ovaries. At age 3⁺, 13 of 19 triploid females had oocytes at various stages of development, including perinucleolar, yolk vesicle and yolk globule stages. In addition, three of these fish had collectively produced 72 mature stage oocytes. Thus, whereas diploid brook trout can produce mature oocytes as two-year-olds, triploids cannot do so until four years of age, with the number of mature oocytes being greatly reduced.     

Changes of vitellogenin and Lipase in captive Sterlet sturgeon <Emphasis Type="Italic">Acipenser ruthenus</Emphasis> females during previtellogenesis to early atresia

Plasma chemistry, lipid metabolism and vitellogenin gene expression of captive Sterlet sturgeon Acipenser ruthenus were studied in different maturity stages. A total of 32 fish were sampled, and maturity stages were identified on the basis of histological criteria and direct observation. Females were classified to four groups: previtellogenic, vitellogenic, post-vitellogenic, and atresia. Blood, gonad and liver tissue samples were taken through non-lethal biopsy. Our results showed that plasma levels of glucose, cholesterol, triacylglycerol, high-density lipoprotein, low-density lipoprotein, very low-density lipoprotein, calcium, phosphorus, alkaline phosphatase activity, albumin and total protein increased during ovarian development and were highest at post-vitellogenic stage. The lowest amounts in atresia stage demonstrate that lipid and energy imbalance was related to reabsorption and digestion of the yolk. These results suggested that the VLDL was the main plasma lipoprotein component of Sterlet. We determined that lipoprotein lipase and hepatic lipase activity increased during vitellogenesis process which suggested the role of lipase enzymes in regulating blood lipid metabolism. RT-PCR analysis indicates that Vitellogenin (VTG) mRNA could be detected both in livers and ovaries of female Sterlet. Throughout the study, the expression level of VTG gene showed an increase both in ovaries and in livers reaching its peak at late vitellogenesis stage. This strongly indicated a relation between VTG mRNA and ovarian development.     

Determinism of the induction of the reproductive cycle in female Eurasian perch, Perca fluviatilis: Identification of environmental cues and permissive factors

In Eurasian perch, Perca fluviatilis, gametogenesis is induced by decrease of both temperature and photoperiod. However, a multiplicity of other factors can display non-permissive or modulating effects on the induction of the reproductive cycle. Consequently, a 9-week study was carried out to determine the environmental cues and permissive factors controlling the induction of the reproductive cycle in this species. A two-level fractional factorial design 2^8-4 was used in order to identify the influent factors from 8 environmental and nutritional ones and their 28 interactions. Tested factors were: (1) amplitude of temperature decrease, (2) time, (3) kinetics and (4) amplitude of photoperiod decrease, (5) initial nutritional state, (6) handling, (7) feeding rate and (8) light spectrum. Increase of gonadosomatic index (GSIi), proportion of female in exogenous vitellogenesis (EVP), plasma 17 β-estradiol (E₂) and cortisol levels, fat consumption and food intake were evaluated.This study showed that the amplitude of the decrease of temperature and the time of photoperiod decrease are the two main environmental cues controlling the induction of the reproductive cycle in Eurasian perch. GSIi was the highest (3.8%) when females were exposed to both low amplitude of temperature decrease and precocious decrease of photophase. It was the lowest (1.1%) when high amplitude of temperature decrease and late decrease of photophase were applied. Handling in association with temperature modulated the broodstock response. 100% of the unhandled fish held at 14 °C were at the exogenous vitellogenesis stage with mean E₂ and basal cortisol levels of 1.6 ng mL^− 1 and 9 ng mL^− 1 respectively. Handled fish and fish held at 6 °C exhibited lower vitellogenesis (40-73%) associated with lower E₂ (0.6-1.1 ng mL^− 1) and higher basal cortisol levels (37-89 ng mL^− 1). No other factor nor interaction displayed a cueing or non permissive effect on the induction of the reproductive cycle in female Eurasian perch. A first schematic model is proposed to explain the factorial determinism of the induction of the reproductive cycle.     

Dynamics of glucose in the haemolymph of female giant freshwater prawn,Macrobrachium rosenbergii,influences reproductive and non‐reproductive moulting cycles

Glucose, when measured in haemolymph, has been found to reflect a useful predictor of energetic investment. This study evaluated the pattern of glucose in the haemolymph, with an attempt to gain a better insight into the role of glucose as nutritional source of ovarian development and energy reserves during reproductive and non‐reproductive moulting cycles. The haemolymph of female giant freshwater prawn, Macrobrachium rosenbergii, was obtained at eight different moulting stages, and levels of glucose were determined using an enzymatic colorimetric glucose‐oxidase method in parallel with a histological examination of ovarian development. Glucose levels were relatively low (0.15 ± 0.02 mg mL^?1) at D₀ stage, an abrupt increase (0.52 ± 0.13 mg mL^?1) during premoult D₁ stage and declined (0.32 ± 0.10 and 0.31 ± 0.09 mg mL^?1) during premoult D₂ and D₃ stages, respectively; thereafter, a slight increase (0.43 ± 0.09 mg mL^?1) occurred at post‐moult A stage. The progression of ovarian growth, marked by an increasing gonadosomatic index (GSI) pattern during the reproductive moulting cycle (C₀–D₃ stages), was directly proportionate to fluctuations in glucose levels. GSI was significantly positively correlated with glucose (R = 0.40; P < 0.05). In contrast, glucose was notably higher at post‐moult A and premoult D₂ stages during non‐reproductive moulting cycle, the period during which glucose is crucial for exoskeletal chitin synthesis. At this particular stage, a negative correlation between body weight and glucose (R = ?0.36; P < 0.05) was observed. The dynamics of glucose in the haemolymph of female M. rosenbergii correlated with ovarian growth, which signify that glucose as nutritional source for vitellogenesis, and affects the body weight of this species.     

Dry or wet? What is the best choice to determine gonadosomatic and hepatosomatic indexes in females of Macrobrachium amazonicum?

The gonadosomatic (GSI) and hepatosomatic (HSI) indexes are widely used for the evaluation of reproductive biology in crustaceans. However, the use of either wet or dry weight for estimating these indexes is not standardized in the literature. Here, we compare the GSI and HSI indexes based on wet and dry samples of females of Macrobrachium amazonicum from populations with different phenotypes (large and small‐size prawns) as a model. In our analysis, for GSI, the application of wet weight tended to overestimate the values recorded in the initial (I‐II) and intermediate (III) stages of ovarian development, and underestimate the results in the final (IV‐V) stages when compared to the use of dry weight. This could be attributed to the reduction in cytoplasmic organelles and water content of the cytosol due to the increase in yolk and lipids in the oocytes throughout ovarian development. For HSI, no significant differences were detected between wet and dry samples. However, the dry HSI computed at all stages of development was overestimated when compared to the use of wet samples, and only the hepatopancreas of females at the final stage (V) showed a significant decrease in water content. Thus, our results indicate that M. amazonicum females maintain constant hepatopancreatic reserves throughout ovarian development. In this way, the use of dry weight is more adequate to estimate GSI and HSI in this species.     

Purification and development of ELISAs for two forms of vitellogenin in Indian walking catfish, <Emphasis Type="Italic">Clarias batrachus</Emphasis> (L.)

Two forms of vitellogenin (Vg: Vg1 and Vg2) were purified from the plasma of estradiol-17β (E₂)-treated Indian walking catfish, Clarias batrachus, by gel filtration and adsorption chromatography. Native Vg1 and Vg2 had apparent molecular masses of 375 and 450 kDa, respectively, and both Vgs resolved into two similar major bands (95 and 67 kDa) in SDS-PAGE under reducing condition. Polyclonal antisera raised against each form of Vg were absorbed with a combination of hypophysectomized male catfish serum proteins and alternate Vg to ensure specificity. Immunological analyses verified the presence of Vg1 and Vg2 in the plasma of female catfish. Homologous ELISAs were developed for Vg1 and Vg2 using their respective harvested antisera, which exhibited the detection limit of 100 ng ml^?1 for Vg1 and 40 ng ml^?1 for Vg2, and low level of cross-reactivity (not parallel to the standard) was found with alternate Vg in each assay. Treatment of male catfish with E₂ induced both Vgs showing a proportionate ratio of Vg1 to Vg2 at 5.6:1. Plasma concentrations of both Vgs measured by ELISAs at different reproductive phases of field collected female catfish increased in accordance with the ovarian development, keeping the proportionate ratio of Vg1 to Vg2 at about 2:1 in fish undergoing vitellogenesis during prespawning period and 1:20 during spawning period, suggesting that Vg1 may be the major Vg to contribute in yolk formation, whereas Vg2, besides its role in yolk formation, may facilitate other physiological functions. The present study, thus, demonstrates the occurrence of two unequally synthesized Vgs in the catfish.     

Effects of low dietary protein level on serum oestradiol,testosterone and sex reversal in rice field eel,Monopterus albus (Zuiew)

We investigated the effects of low dietary protein in isocaloric diets on sex reversal of Monopterus albus by evaluating the oestradiol (E₂) and testosterone (T) concentrations, gonadosomatic index (GSI), sex ratio and gonad structure at the histological level. Fish (9.50 ± 1.50 g average initial weight; n=3 per group) were fed with five practical diets containing 100, 150, 200, 250 or 400 g kg^?1 crude protein to apparent satiation for 15 months. Serum E₂ and T concentrations were determined by radioimmunoassays. E₂ concentrations and GSI significantly increased while T concentrations decreased as the dietary protein level was raised. Fish fed 400 g kg^?1 of dietary protein had significantly higher E₂ concentrations and GSI than those fed with lower dietary protein levels. The T concentrations of fish fed 100 g kg^?1 of dietary protein was significantly higher than that of fish fed higher dietary protein levels. The shift of sex ratio towards more male and intersex fish was observed with decreasing dietary protein levels. Therefore, low dietary protein level may promote sex change from female to male in M. albus. This study provides important information for successful reproductive management and may be exploited for aquaculture of this species.     

西伯利亚鲟不同性别与卵巢发育时期血液性类固醇激素的差异与判别分析

利用电化学发光免疫分析法(ECLIA)测定了人工养殖西伯利亚鲟(Acipenser baerii baerii)不同性别和卵巢发育时期血浆中睾酮(T)和雌二醇(E2)的含量。利用活组织取样的方法,根据性别和卵子颜色及大小将实验鱼分为成熟雄鱼(M)、雌鱼Ⅲ期、Ⅳ期和Ⅴ期共4个组。研究结果表明,西伯利亚鲟血浆T含量雄鱼高于雌鱼(P<0.01),且雄鱼与雌鱼Ⅲ期和Ⅴ期组有极显著性差异(P<0.01),而E2含量除与雌鱼Ⅳ期有极显著性差异外(P<0.01),与雌鱼Ⅲ期和Ⅴ期组无显著性差异(P>0.05),雄鱼表现为T含量和T/E2比值高,而E2含量低。雌鱼随卵巢的发育T和E2含量均表现为"先上升,后下降"的趋势,即Ⅳ期组性类固醇激素的含量明显高于Ⅲ期和Ⅴ期,且差异极显著(P<0.01)。根据血浆中T和E2的含量建立了西伯利亚鲟性别和性腺发育时期的4个判别函数:YM=-11.322+0.043T+0.153E2,YⅢ=-1.390+0.000T+0.061E2,YⅣ=-18.074+0.032T+3.305E2,YⅤ=-2.316+0.785T+0.008E2,经检验总体判别准确率为95%,其中对性别的判别准确率为100%,对雌鱼卵巢发育时期的判别准确率为93%。     

Evidences for involvement of growth hormone and insulin-like growth factor in ovarian development of starry flounder (<Emphasis Type="Italic">Platichthys stellatus</Emphasis>)

Although gonadotrophins are major regulators of ovarian function in teleosts and other vertebrates, accumulating evidence indicates that the growth hormone (GH)-insulin-like growth factor (IGF) axis also plays an important role in fish reproduction. As a first step to understand the physiological role of the GH-IGF system in the ovarian development of starry flounder (Platichthys stellatus), the expression profiles of GH and IGF messenger RNAs (mRNAs) and plasma GH, IGF-I, estradiol-17β (E2), and testosterone (T) levels during the ovarian development were investigated. The developmental stages of ovaries were divided into five stages (II, III, IV, V, and VI) by histological analysis. The hepatosomatic index (HSI) and gonadosomatic index (GSI) values increased and peaked at stage IV and stage V, respectively, and then declined at stage VI. Pituitary GH mRNA levels decreased sharply at stage III and raised to top level at stage VI. The hepatic IGF-I mRNA levels ascended to maximum value at stage V and then declined significantly at stage VI. However, the hepatic IGF-II mRNA levels remained stable and increased significantly at stage VI. In contrast, the ovarian IGF-I mRNA levels increased gradually and peaked at stage VI. The ovarian IGF-II mRNA levels were initially stable and increased significantly at stage V until the top level at stage VI. Consistent with the pituitary GH mRNA levels, plasma GH levels reduced sharply at stage III and remained depressed until stage V and then raised remarkably at stage VI. Plasma IGF-I level peaked at stage V and then declined to initial level. Plasma E2 level peaked at stage IV and then dramatically descended to the basal level. Plasma T level peaked at stage V and then declined significantly back to the basal level. Based on statistical analysis, significant positive correlations between hepatic IGF-I mRNA and GSI, ovarian IGF-II mRNA and hepatic IGF-II mRNA, ovarian IGF-I mRNA and ovarian IGF-II mRNA, and plasma IGF-I and plasma T were observed, respectively. These results suggest that the GH-IGF system may be involved in the ovarian development of starry flounder; GH and IGFs appear to play distinct roles in the regulation of the ovarian development in paracrine/autocrine manners. These findings extend our knowledge of the roles of the GH-IGF axis on reproduction regulation in fish.